• 대한한방소아과학회지
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• 제24권1호
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• pp.126-133
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• 2010

Objectives In this study, we evaluated the therapeutic effects of Gami-Bojungikgitang and Gami-Jwagwieum for bleomycin-induced lung fibrosis in mice. Methods Extracted lyophilization, Gami-Bojungikgitang (96g) and Gami-Jwagwieum (118g) boiled, filtered, depressed, concentrated, and are obtained. They were divided into five groups: normal, group IA; Animal group treated with bleomycin observed on the 21th day, group IB; Animal group treated with bleomycin observed on the 42th day, group IIA; Animal group treated with bleomycin and Gami-Bojungikgitang. Gami-Jwagwieum observed on the 21th day, group IIB; Animal group treated with bleomycin and Gami-Bojungikgitang/Gami-Jwagwieum observed on the 42th day. Mice are used on the 42th day and as a result, bronchoalveolar lavages fluid is obtained. Counting total number of cells, different ratio of macrophage, lymphocyte, and neutrophil are established. Results In animal group treated with bleomycin and Gami-Bojungikgitang, total cell count decreased by 50% in 3 weeks compared to animal group with non-administrated Gami-Bojungikgitang. However, total cell count in 6 weeks increased compared to 3 weeks although total cell count still decreased compared to animal group with non-administrated Gami-Bojungikgitang. In the view of differential cell counts in bronchoalveolar lavages fluid in treatment group on 3 and 6 weeks, neutrophile was a few and lymphocyte decreased. In animal group treated with bleomycin and Gami-Jwagwieum, total cell count decreased by 50% in 3 and 6 weeks compared to animal group with non-administrated Gami-Jwagwieum. In the view of differential cell counts in bronchoalveolar lavages fluid in treatment group on 3 and 6 weeks, lymphocyte also decreased. Conclusions Gami-Bojungikgitang and especially Gami-Jwagwieum for bleomycin-induced lung fibrosis in mice were effective in total cell count and differential cell count.

• 대한한방종양학회지
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• 제5권1호
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• pp.77-101
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• 1999

l. Sojeokbaekchoolsan compound treatment decreased pulmonary fibrosis induced by Bleomycin. 2. At 1 and 2 days after bleomycin treatment, Sojeokbaekchoolsan compound treatment decreased the number of neutrophils in bronchoalveolar lavage than those of bleomycin alone treatment. 3. Sojeokbaekcoolsan compound treatment increased the Fc receptor mediated rosette activity of alveolar macrophage decreased by bleomycin treatment. 4, At 10 days after bleomycin treatment, Sojeokbaekchoolsan compound treatment decreased the lipid peroxidation of lung tissue than those of bleomycin alone treatment. 5. Anti-tumor activity of Sojeokbaekchoolsan and bleomycin compound group was higher than those of bleomycin alone treated group to ascitic tumor caused by Sarcoma-180 tumor cells.

• 대한한방내과학회지
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• 제25권4호
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• pp.93-104
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• 2004

Objectives : The aim was to identify the inhibitory effects of Liriopis Tuber on bleomycin-induced lung fibrosis by analysing the changes of imflammatory cell cytokines and SHI(Semiquantitative Histological Index), Materials and Methods : In this study fibrosis prone C57BL/6J mice were used. Control group was treated with blomycin(0.06mg/0.1 ml) by IT(intratracheal) instillation which is a popular method of inducing lung fibrosis and sample group took Liriopis Tuber water extract(38.0mg/10g body weight) orally for 14 days after IT instillation of blomycin. We measured the total and differential count of WBC, $IFN-{\gamma}$ & IL-4 in mice BALF and SHI(Semiquantitative Histological Index) from lung tissues of mice. BALF and lung tissues of mice were taken 14 days after IT instillation of blomycin. Results : In sample group total WBC count, proportion of neutrophil, SHI and IL-4 significantly(p<0.05) decreased, proportion of macrophage significantly(p<0.05) increased and proportion of lymphocyte, $IFN-{\gamma}$ did not decrease significantly. Conclusions : This study suggests that Liriopis Tuber has an inhibitory effects of pulmonary fibrosis by attenuation of inflammation and Th2 immune response. To determine whether this herbal medicine contribute to cure and prophylaxis of pulmonary fibrosis, further studies on the role of $IFN-{\gamma}$ relating to fibrosis are required.

• 혜화의학회지
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• 제29권1호
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• pp.1-17
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• 2020

Objectives : The objectives of this study is to develop a mouse model similar to Taeeum-type by inducing Lung fibrosis with bleomycin, and to determine adequate concentration of bleomycin. Methods : The subjects were divided into six groups: normal, obesity induced group, and bleomycin administered 0.015U, 0.03U, 0.06U, and 0.09U(U/100g bw) concentrations respectively. Each concentration of bleomycin was dissolved in distilled water, and administered through Intra-Nazal-Trachea injection method. Food intake and body weight were measured at regular time weekly. At the end of the experiment, blood was gathered by cardiac puncture for biochemical examinations, organs were removed for histological examinations, and weigh and mRNA genes was analyzed. Result : Mice administered with bleomycin at 0.015U and 0.03U showed body and fat weight gain, and increased blood total cholesterol, LDL-cholesterol, glucose, and free fatty acid level. Fat related genes also showed higher level than the control group. Obesity was most strongly induced in the mice administered with 0.03U of bleomycin. On the other hand, when bleomycin was administered at concentrations above 0.06U, a model of obesity mouse was not created due to rapid emphysema inflammation and weakness. Conclusions : Mice were most vulnerable to obesity when bleomycin was administered at a concentration of 0.3 to cause liver damage. Bleomycin concentration over 0.06U did not cause obesity-induced mice, due to severe damage in liver.

• Toxicological Research
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• 제26권3호
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• pp.217-222
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• 2010

Many studies have reported that bleomycin, anti-cancer drug, induces pulmonary fibrosis as a side effect. However, few investigations have focused on the dose-response effects of bleomycin on pulmonary fibrosis. Therefore, in the present study, we investigated the effects of different doses of bleomycin in male mice. ICR mice were given 3 consecutive doses of bleomycin: 1, 2, or 4 mg/kg in bleomycin-treated (BT) groups and saline only in vehicle control (VC) groups. The animals were sacrificed at 7 and 24 days postinstillation. The severity of pulmonary fibrosis was evaluated according to inflammatory cell count and lactate dehydrogenase (LDH) activity in the broncho alveolar lavage fluid (BALF), and lung tissues were histologically evaluated after hematoxylin and eosin (H&E), and Masson's trichrome staining. BT groups exhibited changed cellular profiles in BAL fluid compared to the VC group, which had an increased number of total cells, neutrophils, and lymphocytes and a modest increase in the number of macrophages at 7 days post-bleomycin instillation. Moreover, BT groups showed a dose-dependent increase in LDH levels and inflammatory cell counts. However, at 24 days after treatment, collagen deposition, interstitial thickening, and granulomatous lesions were observed in the alveolar spaces in addition to a decrease in inflammatory cells. These results indicate that pulmonary fibrosis induced by 4 mg/kg bleomycin was more severe than that induced by 1 or 2 mg/kg. These data will be utilized in experimental animal models and as basic data to evaluate therapeutic candidates through non-invasive monitoring using the pulmonary fibrosis mouse model established in this study.

• Biomolecules & Therapeutics
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• 제31권4호
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• pp.434-445
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• 2023

We investigated whether FTY-720 might have an effect on bleomycin-induced pulmonary fibrosis through inhibiting TGF-β1 pathway, and up-regulating autophagy. The pulmonary fibrosis was induced by bleomycin. FTY-720 (1 mg/kg) drug was intraperitoneally injected into mice. Histological changes and inflammatory factors were observed, and EMT and autophagy protein markers were studied by immunohistochemistry and immunofluorescence. The effects of bleomycin on MLE-12 cells were detected by MTT assay and flow cytometry, and the related molecular mechanisms were studied by Western Blot. FTY-720 considerably attenuated bleomycin-induced disorganization of alveolar tissue, extracellular collagen deposition, and α-SMA and E-cadherin levels in mice. The levels of IL-1β, TNF-α, and IL-6 cytokines were attenuated in bronchoalveolar lavage fluid, as well as protein content and leukocyte count. COL1A1 and MMP9 protein expressions in lung tissue were significantly reduced. Additionally, FTY-720 treatment effectively inhibited the expressions of key proteins in TGF-β1/TAK1/P38MAPK pathway and regulated autophagy proteins. Similar results were additionally found in cellular assays with mouse alveolar epithelial cells. Our study provides proof for a new mechanism for FTY-720 to suppress pulmonary fibrosis. FTY-720 is also a target for treating pulmonary fibrosis.

• Tuberculosis and Respiratory Diseases
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• 제56권1호
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• pp.51-66
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• 2004

연구배경 : 폐 섬유화증의 진행에는 세포외 간질의 대사에 관여하는 gelatinase가 중요한 역할을 하는 것으로 알려져 있으며, gelatinase의 작용은 사이토카인을 비롯한 여러 가지 요소에 의해 조절되는 것으로 알려져 있다. 지금까지 $interferon-{\gamma}$($IFN-{\gamma}$)는 폐섬유화를 억제하는 것으로 알려져 있지만 gelatinase의 작용에 $IFN-{\gamma}$가 미치는 영향은 잘 밝혀져 있지 않다. 이에 저자들은 bleomycin 유도 백서 폐 섬유화 모델에서 $IFN-{\gamma}$가 폐 섬유화에 미치는 영향과MMP-2, -9과 이의 길항제인 TIMP-1, -2 그리고 Th-2 사이토카인의 변화에 미치는 영향을 조사함으로써, $IFN-{\gamma}$가 폐 섬유화증에 미치는 영향과 기전을 연구하고자 하였다. 재료 및 방법 : Sprague-Dawley계의 수컷 흰쥐를 정상 대조군, bleomycin 군, bleomycin+$IFN-{\gamma}$ 군의 세 군으로 나누어, bleomycin 군 과 bleomycin+$IFN-{\gamma}$ 군은 bleomycin sulfate를 생리식염수에 섞어 기관 내로 주입하였고(6 U/kg), bleomycin+$IFN-{\gamma}$ 군은 $IFN-{\gamma}$를 14일간 근육주사 하였다($2{\times}105U/kg$). 실험동물은 3, 7, 14, 28일에 폐를 얻어 hematoxylineosin 염색과 Masson's trichrome 염색을 하여 폐의 염증 반응과 섬유화 정도를 관찰하였고, 폐 조직 내의 hydroxyproline 함량, MMP-2, -9, TIMP-1, -2에 대한 Western blot, zymography와 reverse zymography, IL-4와 IL-13에 대한 ELISA검사를 시행하였다. 결 과 : 1. 폐 염증은 bleomycin 투여 7일째에는 $IFN-{\gamma}$를 투여한 경우 더 심하게 일어났으나(bleomycin 군 : bleomycin+$IFN-{\gamma}$ 군=$2.08{\pm}0.15:2.74{\pm}0.29$, P<0.05), 폐 섬유화는 bleomycin 투여 후 28일째 $IFN-{\gamma}$ 투여에 의해 유의하게 감소되었다(bleomycin 군 : bleomycin+$IFN-{\gamma}$ 군=$3.94{\pm}0.43:2.46{\pm}0.13$, P<0.05). 2. 폐 내 hydroxyproline 함량은 bleomycin 투여 28일 후 $IFN-{\gamma}$ 투여에 의해 유의하게 감소되었다(bleomycin 군 : bleomycin+$IFN-{\gamma}$ 군=$294.04{\pm}31.73{\mu}g/g:194.92{\pm}15.51{\mu}g/g$, P<0.05). 3. Bleomycin 투여에 의해 MMP-2 단백질의 양이 증가되는 소견이 Western blot에서 관찰되었는데 MMP-2의 증가는 bleomycin 투여 14일에 최고에 이르렀으며, $IFN-{\gamma}$의 투여에 의해 bleomycin에 의한 MMP-2의 증가는 억제되었으나 대조군 수준으로 감소하지는 않았다. 4. Zymography 검사에서 bleomycin 투여 3일째에는 활성화된 형태의 MMP-2가 $IFN-{\gamma}$에 의해 유의하게 증가되었으나(bleomycin 군 : bleomycin+$IFN-{\gamma}$ 군=$209.63{\pm}7.60%:407.66{\pm}85.34%$, P<0.05), 투여 14일 후에서는 $IFN-{\gamma}$에 의해 활성화된 상태의 MMP-2가 유의하게 감소되었다(bleomycin 군 : bleomycin+$IFN-{\gamma}$ 군=$159.36{\pm}20.93%:97.23{\pm}12.50%$, P<0.05). 5. Bleomycin을 투여한 후 bleomycin 군과 bleomycin+$IFN-{\gamma}$ 군 모두에서 IL-4의 양이 감소되었으나, 두 군간 차이는 통계적으로 유의하지는 않았고, IL-13은 별 다른 차이가 없었다. 결 론 : Bleomycin에 의한 백서 폐 섬유화 모델에서 $IFN-{\gamma}$는 초기 폐 염증을 증가시키지만 후기 폐 섬유화는 억제시킨다는 것을 알 수 있었다. $IFN-{\gamma}$가 폐 섬유화를 억제하는 것은 MMP-2의 활성화를 억제하기 때문인 것으로 생각한다. 또한 $IFN-{\gamma}$에 의한 MMP-2의 기능 억제는 Th-2 사이토카인을 억제하여 이루어지는 것 같지는 않았다. 본 논문을 기초로 향후 $IFN-{\gamma}$가 MMP-2의 활성화를 조절하는 기전과 MMP-2의 활성화가 폐섬유화에 미치는 영향에 대한 연구가 필요할 것으로 생각한다.

• 대한한방내과학회지
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• 제25권2호
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• pp.298-306
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• 2004

Objective : The main purpose of this study was to determine if Rehmanniae Radix has significant effects on lung fibrosis. Materials and Methods : C57BL/6J mice were devided into three groups. These were the normal group, which were not treated, the control group, given Intratracheal instillation(IT) of Bleomycin, the sample group, given IT of bleomycin and water-extracted Rehmanniae Radix. Animals were sacrificed 14 days after IT treatment. Lung fibrosis was evaluated by analysis of bronchoalveolar larvage(BAL) total WBC, percentage of macrophage, lymphocyte and neutrophil. This was done histologically by semiquantitative histological index(SHT) of lung tissue. Results : The sample group in coparison with control group showed a decrease in the BAL, total WBC, lower percentage of lymphocyte and neutrophil(p<0.05) and correspondingly a lower percentage of macrophage(p<0.01). The Sample group showed a significant decrease of collagen accumulation with respect to the control group in SHI of lung tissue(p<0.01). INF-${\gamma}$ and IL-4 levels in BALF of mice significantly decreased in the control group(p<0.05). Conclusions : Results suggest that Rehmanniae Radix has an anti-imflamatory effect and anti-fibrotic effect on the lungs through decrease of IL-4 and total WBC count for not only macrophage, but also lymphocyte and neutrophil. The degradation of INF-${\gamma}$ calls for research beyond the scope of this study.

• 사상체질의학회지
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• 제28권2호
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• pp.147-162
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• 2016

Objectives The objective of this study is to develop a taeeumin animal-experimental model induced lung fibrosis with Bleomycin and evaluate the effect on obesity in this animal-experimental model.Methods The subjects were divided into 3 groups : normal group, high fat diet(HFD) control group, and HFD group administered with bleomycin(n=10 per group). To develop taeeumin animal-experimental model with reduced respiratory metabolism, 8-week-old C57BL/6 mice were administered with 0.03ml solution of bleomycin 1U/ml dissolved in distilled water, intratracheal(IT), once. Then, the HFD control group and the experimental group were fed with high fat diet for 6 weeks. Airway hyperresponsiveness(AHR) to methacholine was measured at the 1st and 3rd week after bleomycin was administered. Food intake and body weight were measured at regular time weekly. After the final experiment, blood was gathered by cardiac puncture for bloodchemical examination and organs(liver, fatty tissue) were remoed, weighted, and mRNA was analyzed.Results and Conclusions Through the experiment, it was found that Bleomycin induced Taeeumin animal-experimental models have leptin resistace. In the experimental group administered with Bleomycin, fatty acid synthesizing gene expression increased and energy metabolizing gene expression decreased. As mRNA expression of adiponectin decreased, it was found that Taeeuim animal-experimental model is susceptible to metabolic syndrome and cardiovascular diseases.

• IMMUNE NETWORK
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• 제15권3호
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• pp.142-149
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• 2015

Lung fibrosis is a life-threatening disease caused by overt or insidious inflammatory responses. However, the mechanism of tissue injury-induced inflammation and subsequent fibrogenesis remains unclear. Recently, we and other groups reported that Th17 responses play a role in amplification of the inflammatory phase in a murine model induced by bleomycin (BLM). Osteopontin (OPN) is a cytokine and extracellular-matrix-associated signaling molecule. However, whether tissue injury causes inflammation and consequent fibrosis through OPN should be determined. In this study, we observed that BLM-induced lung inflammation and subsequent fibrosis was ameliorated in OPNdeficient mice. OPN was expressed ubiquitously in the lung parenchymal and bone-marrow-derived components and OPN from both components contributed to pathogenesis following BLM intratracheal instillation. Th17 differentiation of $CD4^+$ ${\alpha}{\beta}$ T cells and IL-17-producing ${\gamma}{\delta}$ T cells was significantly reduced in OPN-deficient mice compared to WT mice. In addition, Th1 differentiation of $CD4^+$ ${\alpha}{\beta}$ T cells and the percentage of IFN-$\gamma$-producing ${\gamma}{\delta}$ T cells increased. T helper cell differentiation in vitro revealed that OPN was preferentially upregulated in $CD4^+$ T cells under Th17 differentiation conditions. OPN expressed in both parenchymal and bone marrow cell components and contributed to BLM-induced lung inflammation and fibrosis by affecting the ratio of pathogenic IL-17/protective IFN-$\gamma$ T cells.